(Hydrazido)(amino)thiophene compounds

ABSTRACT

Compounds represented by Formula (I): 
                         
or a pharmaceutically acceptable salt or N-oxide thereof, wherein A and J are defined herein, are useful in the treatment of tumors and cancers such as mastocytosis/mast cell leukemia, gastrointestinal stromal tumors (GIST), germ cell tumors, small cell lung carcinoma (SCLC), sinonasal natural killer/T-cell lymphoma, testicular cancer (seminoma), thyroid carcinoma, malignant melanoma, ovarian carcinoma, adenoid cystic carcinoma, acute myelogenous leukemia (AML), breast carcinoma, pediatric T-cell acute lymphoblastic leukemia, neuroblastoma, mast cell leukemia, angiosarcoma, anaplastic large cell lymphoma, endometrial carcinoma, and prostate carcinoma.

This application claims the benefit of U.S. Patent Application No.60/610,932 filed 17 Sep. 2004.

BACKGROUND OF THE INVENTION

The present invention is directed to disubstituted thiophenes. Inparticular, the present invention is directed to(2-hydrazido)(3-amino)thiophenes and (2-alkoxamido)(3-amino)thiophenesthat are inhibitors of c-Kit proto-oncogene (also known as Kit, CD-117,stem cell factor receptor, mast cell growth factor receptor).

The c-Kit proto-oncogene is believed to be important in embryogenesis,melanogenesis, hematopoiesis, and the pathogenesis of mastocytosis,gastrointestinal tumors, and other solid tumors, as well as certainleukemias, including AML. Accordingly, it would be desirable to developnovel compounds that are inhibitors of the c-Kit receptor.

Many of the current treatment regimes for hyperproliferative disorders(cancer) utilize compounds that inhibit DNA synthesis. Such compounds'mechanism of operation is to be toxic to cells, particularly to rapidlydividing tumor cells. Thus, their broad toxicity can be a problem to thesubject patient. However, other approaches to anti-cancer agents thatact other than by the inhibition of DNA synthesis have been explored totry to enhance the selectivity of the anti-cancer action and therebyreduce adverse side-effects.

It is known that a cell may become cancerous by virtue of thetransformation of a portion of its DNA into an oncogene (i.e. a genewhich, on activation, leads to the formation of malignant tumor cells).Many oncogenes encode proteins that are aberrant protein-tyrosinekinases capable of causing cell transformation. By a different route,the overexpression of a normal proto-oncogenic tyrosine kinase can alsoresult in proliferative disorders, sometimes resulting in a malignantphenotype. Alternatively, co-expression of a receptor tyrosine kinaseand its cognate ligand within the same cell type may also lead tomalignant transformation.

Receptor tyrosine kinases are large enzymes which span the cell membraneand possess i) an extracellular binding domain for growth factors suchas KIT ligand (also known as stem cell factor (SCF), Steel factor (SLF)or mast cell growth factor (MGF)), ii) a transmembrane domain, and iii)an intracellular portion which functions as a kinase to phosphorylatespecific tyrosine residues in proteins. Binding of KIT ligand to KITtyrosine kinase results in receptor homodimerization, the activation ofKIT tyrosine kinase activity, and the subsequent phosphorylation of avariety of protein substrates, many of which are effectors ofintracellular signal transduction, These events can lead to enhancedcell proliferation or promote enhanced cell survival. With some receptorkinases, receptor heterodimerization can also occur.

It is known that such kinases are frequently aberrantly expressed incommon human cancers such as breast cancer, head and neck cancers,gastrointestinal cancer such as colon, rectal or stomach cancer,leukemia, and ovarian, bronchial, lung or pancreatic cancer. Kit kinaseexpression has been documented in a wide variety of human malignanciessuch as mastocytosis/mast cell leukemia, gastrointestinal stromal tumors(GIST), small cell lung carcinoma (SCLC), sinonasal naturalkiller/T-cell lymphoma, testicular cancer (seminoma), thyroid carcinoma,malignant melanoma, ovarian carcinoma, adenoid cystic carcinoma, acutemyelogenous leukemia (AML), breast carcinoma, pediatric T-cell acutelymphoblastic leukemia, angiosarcoma, anaplastic large cell lymphoma,endometrial carcinoma, and prostate carcinoma. The kinase activity ofKIT has been implicated in the pathophysiology of several of these—andadditional tumors—including breast carcinoma, SCLC, GIST, germ celltumors, mast cell leukemia, neuroblastoma, AML, melanoma and ovariancarcinoma.

Several mechanisms of KIT activation in tumor cells have been reported,including activating mutations, autocrine and paracrine activation ofthe receptor kinase by its ligand, loss of protein-tyrosine phosphataseactivity, and cross activation by other kinases. The transformingmechanisms initiated by the activating mutations are thought to includedimer formation and increased intrinsic activity of the kinase domain,both of which result in constitutive ligand-independent kinaseactivation, and possibly altered substrate specificity. More than thirtyactivating mutations of the Kit protein have been associated with highlymalignant tumors in humans.

Accordingly, it has been recognized that inhibitors of receptor tyrosinekinases are useful as selective inhibitors of the growth of mammaliancancer cells. For example, Gleevec™ (also known as imatinib mesylate, orSTI571), a 2-phenylpyrimidine tyrosine kinase inhibitor that inhibitsthe kinase activity of the BCR-ABL fusion gene product, was recentlyapproved by the U.S. Food and Drug Administration for the treatment ofCML. Gleevec™, in addition to inhibiting BCR-ABL kinase, also inhibitsthe KIT kinase and PDGF receptor kinase, although it is not effectiveagainst all mutant isoforms of the KIT kinase. Kit ligand-stimulatedgrowth of MO7e human leukemia cells is inhibited by Gleevec™, which alsoinduces apoptosis under these conditions. By contrast, GM-CSF stimulatedgrowth of MO7e human leukemia cells is not affected by Gleevec™.Further, in recent clinical studies using Gleevec™ to treat patientswith GIST, a disease in which KIT kinase is involved in transformationof the cells, many of the patients showed marked improvement.

These studies demonstrate how KIT kinase inhibitors can treat tumorswhose growth is dependent on KIT kinase activity. Other kinaseinhibitors show even greater kinase selectivity. For example, the4-anilinoquinazoline compound Tarceva™ inhibits only EGF receptor kinasewith high potency, although it can inhibit the signal transduction ofother receptor kinases, probably by virtue of the fact that thesereceptors heterodimerize with EGF receptor.

Although anti-cancer compounds such as those described above make asignificant contribution to the art, there is a continuing need forimproved anti-cancer pharmaceuticals, and it would be desirable todevelop new compounds with better selectivity or potency, or withreduced toxicity or side effects.

International Patent Publication Nos. WO00/71510, WO01/19798, andWO01/38309 describe factor Xa inhibitors.

International Patent Publication No. WO02/066470 describes substitutedalkylamine derivatives. International Patent Publication No. WO02/068406describes substituted amine derivatives. International PatentPublication No. WO93/07751 describes fungicides for the control oftake-all diseases of plants. U.S. Patent Application Publication No.2003/0195231 describes compounds exhibiting thrombopoietin receptoragonism.

International Patent Publication No. WO03/04042 describes alpha-helixmimicry by a class of organic molecules. International PatentPublication No. WO98/49152 describes protease inhibitors. InternationalPatent Publication No. WO01/96323 describes serine protease inhibitors.

International Patent Publication No. WO00/62778 describes cyclic proteintyrosine kinase inhibitors.

International Patent Publication No. WO03/006454 describes fungicidalcompounds. International Patent Publication No. WO01/23382 describespharmaceutically active sulfonyl hydrazide derivatives. InternationalPatent Publication No. WO04/033433 describes pyrazolone compounds andthrombopoietin receptor activators. U.S. Patent Application PublicationNo. 2004/0063765 describes selective inhibitors of bacterial pathogens.

International Patent Publication No. WO02/085908 describes folatemimetics and folate-receptor binding conjugates thereof.

SUMMARY OF THE INVENTION

Compounds represented by Formula (I):

or a pharmaceutically acceptable salt or N-oxide thereof are useful inthe treatment of tumors and cancers such as mastocytosis/mast cellleukemia, gastrointestinal stromal tumors (GIST), germ cell tumors,small cell lung carcinoma (SCLC), sinonasal natural killer/T-celllymphoma, testicular cancer (seminoma), thyroid carcinoma, malignantmelanoma, ovarian carcinoma, adenoid cystic carcinoma, acute myelogenousleukemia (AML), breast carcinoma, pediatric T-cell acute lymphoblasticleukemia, neuroblastoma, mast cell leukemia, angiosarcoma, anaplasticlarge cell lymphoma, endometrial carcinoma, and prostate carcinoma.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to a compound represented by Formula(I):

or a pharmaceutically acceptable salt or N-oxide thereof, wherein:

J is —NR¹R² or —OR¹;

A is aryl, heteroaryl, cycloC₃₋₁₀alkyl, heterocyclyl, cycloC₃₋₁₀alkenyl,or heterocycloalkenyl, each of which is optionally substituted by 1-5independent R⁴ substituents;

R¹ is C₁₋₆alkyl, aryl, heteroaryl, cycloC₃₋₁₀alkyl, heterocyclyl,cycloC₃₋₁₀alkenyl, heterocycloalkenyl, C₂₋₆alkenyl, C₂₋₆alkynyl, each ofwhich is optionally substituted by 1-5 independent R⁴¹ substituents;

R² is C₀₋₆alkyl, aryl, heteroaryl, cycloC₃₋₁₀alkyl, heterocyclyl,cycloC₃₋₁₀alkenyl, heterocycloalkenyl, C₂₋₆alkenyl, C₂₋₆alkynyl, each ofwhich is optionally substituted by 1-5 independent R⁴² substituents;

or R¹ and R², taken together with the N atom to which they are attached,form a heterocyclyl, heterocylcoalkenyl, or heteroaryl, each of which isoptionally substituted with 1-4 independent R⁴¹ substituents; providedR¹ and R² taken together with the N atom to which they are attached donot form a pyrazolyl;

R³ is C₀₋₆alkyl, halogen, C₁₋₆alkyl, or haloalkyl;

R⁴, R⁴¹, and R⁴² each independently is C₀₋₆alkyl, cycloC₃₋₁₀alkyl, oxo,halogen, haloalkyl, cyanoC₀₋₆alkyl, nitroC₀₋₆alkyl, hydroxyC₀₋₆alkyl,C₀₋₆alkylaminoC₀₋₆alkyl, aminoC₁₋₆alkylamino, acylaminoC₀₋₆alkylamino,acylC₀₋₆alkyl, substituted acyl, guanidinoC₀₋₆alkyl,hydroxyiminoC₀₋₆alkyl, acylaminoC₀₋₆alkyl, substituted acylamino,acyloxyC₀₋₆alkyl, substituted acyloxy, arC₀₋₆alkyl, substitutedarC₀₋₆alkyl, heteroarylC₀₋₆alkyl, substituted heteroarylC₀₋₆alkyl,heterocyclylC₀₋₆alkyl, cyanoaminoC₀₋₆alkyl, C₀₋₆alkylhydrazino,heterocyclylamino, arC₀₋₆alkylhydrazino, alkylsulfonylC₀₋₆alkyl,arC₀₋₆alkylsulfonylC₀₋₆alkyl, alkylsulfinylC₀₋₆alkyl,alkylsulfonamidoC₀₋₆alkyl, arC₀₋₆alkylsulfonamidoC₀₋₆alkyl,aminoC₀₋₆alkylsulfonyl, C₀₋₆alkylaminosulfonyl, acylC₁₋₆alkylsulfonyl,heterocyclylsulfonyl, aminoC₀₋₆alkylsulfinyl, acylC₁₋₆alkylsulfinyl,silyl, siloxy, alkenoxy, alkynoxy, C₂₋₆alkenyl, acylC₂₋₆alkenyl,C₂₋₆alkynyl, acylC₂₋₆alkynyl, hydroxyC₂₋₆alkynyl, aminoC₂₋₆alkynyl,C₁₋₆alkoxyC₀₋₆alkyl, C₁₋₆alkylthioC₀₋₆alkyl, hydroxyC₁₋₆alkoxyC₀₋₆alkyl,hydroxyC₁₋₆alkylthioC₀₋₆alkyl, acylC₁₋₆alkoxyC₀₋₆alkyl,acylC₁₋₆alkylthioC₀₋₆alkyl, C₀₋₆alkylaminoC₁₋₆alkoxyC₀₋₆alkyl,C₀₋₆alkylaminoC₁₋₆alkylthioC₀₋₆alkyl, acylaminoC₁₋₆alkoxyC₀₋₆alkyl,acylaminoC₁₋₆alkylthioC₀₋₆alkyl, arC₀₋₆alkylaminoC₀₋₆alkyl,arC₀₋₆alkylthioC₀₋₆alkyl, arC₀₋₆alkoxyC₀₋₆alkyl, arC₀₋₆alkylamino,arC₀₋₆alkylaminoC₀₋₆alkyl, arC₀₋₆alkylthio, substituted arC₀₋₆alkoxy,substituted arC₀₋₆alkylthio,or substituted arC₀₋₆alkoxy; and

provided that the compound is notN′-(tert-butyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazideorN-morpholin-4-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide.

In one aspect, the present invention is directed to a compoundrepresented by Formula (I), or a pharmaceutically acceptable salt orN-oxide thereof, wherein J is —NR¹R²; and the other variables are asdescribed above for Formula (I).

In an embodiment of this one aspect, the present invention is directedto a compound represented by Formula (I), or a pharmaceuticallyacceptable salt or N-oxide thereof, wherein J is —NR¹R²; R³ is hydrogen;and the other variables are as described above for Formula (I).

In another embodiment of this one aspect, the present invention isdirected to a compound represented by Formula (I), or a pharmaceuticallyacceptable salt or N-oxide thereof, wherein J is —NR¹R²; A isheteroaryl; and the other variables are as described above for Formula(I).

In yet another embodiment of this one aspect, the present invention isdirected to a compound represented by Formula (I), or a pharmaceuticallyacceptable salt or N-oxide thereof, wherein J is —NR¹R²; A is

and the other variables are as described above for Formula (I).

In a second aspect, the present invention is directed to a compoundrepresented by Formula (I), or a pharmaceutically acceptable salt orN-oxide thereof, wherein J is —OR¹; and the other variables are asdescribed above for Formula (I).

In an embodiment of this second aspect, the present invention isdirected to a compound represented by Formula (I), or a pharmaceuticallyacceptable salt or N-oxide thereof, wherein J is —OR¹; wherein R³ ishydrogen; and the other variables are as described above for Formula(I).

In another embodiment of this second aspect, the present invention isdirected to a compound represented by Formula (I), or a pharmaceuticallyacceptable salt or N-oxide thereof, wherein J is —OR¹; A is heteroaryl;and the other variables are as described above for Formula (I).

In yet another embodiment of this second aspect, the present inventionis directed to a compound represented by Formula (I), or apharmaceutically acceptable salt or N-oxide thereof, wherein J is —OR¹;A is

and the other variables are as described above for Formula (I).

The present invention includes the following compounds:

-   -   N′-(4-chlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N-(tert-butoxy)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;    -   N-piperidin-1-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;    -   N′-(4-bromophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N-(4-methylpiperazin-1-yl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;    -   N′-(3,4-dichlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N-phenoxy-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;    -   N′-methyl-N′-phenyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N′-(3-chlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N′-phenyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N′-(4-cyanophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   3-[(quinolin-4-ylmethyl)amino]-N′-[4-(trifluoromethyl)phenyl]thiophene-2-carbohydrazide;    -   N′-(4-methoxyphenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   3-[(quinolin-4-ylmethyl)amino]-N′-[3-(trifluoromethyl)phenyl]thiophene-2-carbohydrazide;    -   N′-(4-fluorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;    -   N-2,3-dihydro-1H-indol-1-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide    -   or a pharmaceutically acceptable salt, or N-oxide, thereof.

The present invention is also directed to a method of treatinghyperproliferative disorders, including breast cancer, head cancer, orneck cancer, gastrointestinal cancer, leukemia, ovarian, bronchial,lung, or pancreatic cancer, sinonasal natural killer/T-cell lymphoma,testicular cancer (seminoma), thyroid carcinoma, malignant melanoma,adenoid cystic carcinoma, angiosarcoma, anaplastic large cell lymphoma,endometrial carcinoma, or prostate carcinoma, by administering aneffective amount of a compound represented by Formula (I), or apharmaceutically acceptable salt thereof.

As used herein, “C₀₋₆alkyl” is used to mean an alkyl having 0-6carbons—that is, 0, 1, 2, 3, 4, 5, or 6 carbons in a straight orbranched configuration. An alkyl having no carbon is hydrogen when thealkyl is a terminal group. An alkyl having no carbon is a direct bondwhen the alkyl is a bridging (connecting) group.

As used herein unless otherwise specified, “alkyl”, “alkenyl”, and“alkynyl” includes straight or branched configurations. Lower alkyls,alkenyls, and alkynyls have 1-6 carbons. Higher alkyls, alkenyls, andalkynyls have more than 6 carbons.

As used herein unless otherwise specified, “halogen” is fluorine,chlorine, bromine or iodine.

As used herein unless otherwise specified, “substituted” is used to meanhaving 1-5 independent C₀₋₆alkyl, halogen, nitro, cyano, haloalkyl,C₀₋₆alkoxy, C₀₋₆alkylthio, or C₀₋₆alkylamino substituents

As used herein unless otherwise specified, “haloalkyl” includes alkylgroups substituted with one or more halogens, for example, chloromethyl,2-bromoethyl, 3-iodopropyl, trifluoromethyl, perfluoropropyl,8-chlorononyl, and the like.

As used herein unless otherwise specified, the terms “aryl” and “ar” arewell known to chemists and include, for example, phenyl and naphthyl, aswell as phenyl with one or more short alkyl groups (tolyl, xylyl,mesityl, cumenyl, di(t-butyl)phenyl). Phenyl, naphthyl, tolyl, and xylylare preferred. “Substituted aryl” is an aryl substituted with suitablesubstituents such as, for example, acyl, substituted acyl, N-protectedpiperazinylsulfonyl, piperazinylsulfonyl,N—C₁₋₆alkylpiperazinylsulfonyl, hydroxyC₁₋₆alkyl, heterocyclyl, halogen,nitro, amino, C₁₋₆alkylamino, cyano, or C₁₋₆alkoxy.

As used herein unless otherwise specified, the term “cycloalkyl” is wellknown to chemists and includes cyclic aliphatic ring structures,optionally substituted with alkyl, hydroxyl, oxo, and halo, such ascyclopropyl, methylcyclopropyl, cyclobutyl, cyclopentyl,2-hydroxycyclopentyl, cyclopentanonyl, cyclohexyl, 4-chlorocyclohexyl,cycloheptyl, cyclooctyl, and the like.

As used herein unless otherwise specified, the term “cycloalkyl” is wellknown to chemists and includes cyclic aliphatic ring structures havingat least one ethylenic bond, optionally substituted with alkyl,hydroxyl, oxo, and halo, for example, methylcyclopropenyl,trifluoromethylcyclopropenyl, cyclopentenyl, cyclohexenonyl,cyclohexenyl, 1,4-cyclohexadienyl, and the like.

As used herein unless otherwise specified, “heterocyclyl” is well knownto chemists and includes saturated or partially saturated, mono orpolycyclic heterocyclic groups containing at least one N, S or Ohetero-ring atom such as, for example, tetrahydrofuranyl,tetrahydrofuryl, pyrrolidinyl, piperidinyl, tetrahydropyranyl,thiolanyl, morpholinyl, piperazinyl, homopiperazinyl, dioxolanyl,dioxanyl, indolinyl, or chromanyl and the like. Such heterocyclyls canbe suitably substituted with lower alkyl or oxo substituents.

As used herein unless otherwise specified, “heteroaryl” is well known tochemists and includes partially saturated, mono or polycyclicheterocyclic groups containing at least one N, S or O hetero-ring atomsuch as, for example, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl,pyridazinyl, triazolyl, tetrazolyl, pyrrolidinyl, indolyl, indolinyl,isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl,imidazopyridyl, indazolyl, benzotriazolyl, tetrazolo-pyridazinyl,pyranyl, furyl, thienyl, oxazolyl, isoxazolyl, oxazolyl, benzofuranyl,benzoxazolyl, benzoxadiazolyl, thiazolyl, thiadiazolyl, thiazolidinyl,benzothiazolyl, benzothiadiazolyl, benzofuranyl, or benzodioxyl,imidazolyl, pyrrolyl, oxadiazolyl, quinolyl, benzotriazolyl, orbenzothienyl and the like. Such heterocyclyls can be suitablysubstituted with lower alkyl or oxo substituents.

As used herein unless otherwise specified, “heterocycloalkenyl” includesmono or polycyclic heterocyclic groups having at least one ethylenicbond and containing at least one N, S or O hetero-ring atom such as, forexample, dihydropyranyl, dihydrofuran, pyrrolinyl or the like. Suchheterocycloalkenyls can be suitably substituted with lower alkyl or oxosubstituents.

As used herein unless otherwise specified, “acyl” includes for example,carboxy, esterified carboxy, carbamoyl, lower alkylcarbamoyl, loweralkanoyl, aroyl, heterocyclylcarbonyl, and the like. Esterified carboxyincludes substituted or unsubstituted lower alkoxycarbonyl such asmethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl,t-butoxycarbonyl, hexyloxycarbonyl, 2-iodoethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, dimethylaminopropoxycarbonyl,dimethylaminoethoxycarbonyl; substituted or unsubstitutedaryloxycarbonyl such as phenoxycarbonyl, 4-nitrophenoxycarbonyl,2-naphthyloxycarbonyl; substituted or unsubstitutedar(lower)alkoxycarbonyl such as benzyloxycarbonyl, phenethyloxycarbonyl,benzhydryloxycarbonyl, 4-nitrobenzyloxycarbonyl,3-methoxy-4-nitrobenzyloxycarbonyl; and N-containingheterocyclyloxycarbonyl such as N-methylpiperidyloxycarbonyl and thelike.

As used herein unless otherwise specified, “C₀₋₆alkylhydrazino” may be2-mono or 2,2-di(C₀₋₆alkyl)hydrazino such as 2-methylhydrazino,2,2-dimethylhydrazino, 2-ethylhydrazino, hydrazine,2,2-diethylhydrazino, or the like.

As used herein unless otherwise specified, alkylamino such as“C₁₋₆alkylamino” may be mono or dialkylamino such as methylamino,dimethylamino, N-methylethylamino or the like. Similarly, other aminogroups such as acylamino are understood to include a C₀₋₆alkyl at theunspecified amino bond site (one being to the acyl, the second forming aconnection to the core structure, and the third unspecified).

As used herein unless otherwise specified, “arC₀₋6alkylamino” may bemono or disubstitutedamino such as anilino, benzylamino,N-methylanilino, N-benzylmethylamino or the like.

As used herein unless otherwise specified, “silyl” includes alkyl andaryl substituted silyl groups such as, for example, triethylsilyl,t-butyldiphenylsilyl, or the like.

As used herein unless otherwise specified, “siloxy” includes alkyl andaryl substituted silyloxy groups such as, for example, triethylsilyloxy,t-butyldiphenylsilyloxy, or the like.

As used herein unless otherwise specified, “sulfonyloxy” includessulfonyloxy groups substituted with aryl, substituted aryl, or alkylsuch as, for example, benzenesulfonyl, tosyl, mesyl or the like.

As used herein unless otherwise specified, “heterocyclylamino” includesunsaturated, mono or polycyclic heterocyclic groups containing at leastone N-ring atom which is attached to an amino group such as, forexample, 1-aminopiperidine, 1-aminomorpholine,1-amino-4-methylpiperazine or the like.

As used herein unless otherwise specified (for example, by a dashmarking the point of attachment), chemical group names comprised ofmultiple chemical terms are used according to standard chemicalconvention, wherein each term modifies the following term and whereinthe rightmost term forms a covalent bond with the structure to which thesubstituent is attached. For example, aralkylamino includes benzylaminoand phenethylamino attached through the amino nitrogen, but nottoluidino or N-methylanilino groups.

Formula (I) is shown without a definitive stereochemistry at certainpositions. The present invention includes all stereoisomers of Formula(I) and pharmaceutically acceptable salts thereof. Further, mixtures ofstereoisomers as well as isolated specific stereoisomers are alsoincluded. During the course of the synthetic procedures used to preparesuch compounds, or in using racemization or epimerization proceduresknown to those skilled in the art, the products of such procedures canbe a mixture of stereoisomers.

The invention also encompasses a pharmaceutical composition that iscomprised of a compound of Formula (I) in combination with apharmaceutically acceptable carrier.

Preferably, the composition is comprised of a pharmaceuticallyacceptable carrier and a non-toxic therapeutically effective amount of acompound of Formula (I) as described above (or a pharmaceuticallyacceptable salt or N-oxide thereof).

Moreover, within this preferred embodiment, the invention encompasses apharmaceutical composition for the treatment of disease by theinhibition of the c-Kit kinase, which may be a wild-type or mutant formof the protein, comprising a pharmaceutically acceptable carrier and anon-toxic therapeutically effective amount of compound of Formula (I) asdescribed above (or a pharmaceutically acceptable salt or N-oxidethereof).

The compounds and compositions of the present invention are effectivefor treating mammals such as, for example, humans.

The term “pharmaceutically acceptable salts” refers to salts preparedfrom pharmaceutically acceptable non-toxic bases or acids. When thecompound of the present invention is acidic, its corresponding salt canbe conveniently prepared from pharmaceutically acceptable non-toxicbases, including inorganic bases and organic bases. Salts derived fromsuch inorganic bases include aluminum, ammonium, calcium, copper (ic andous), ferric, ferrous, lithium, magnesium, manganese (ic and ous),potassium, sodium, zinc and the like salts. Particularly preferred arethe ammonium, calcium, magnesium, potassium and sodium salts. Saltsderived from pharmaceutically acceptable organic non-toxic bases includesalts of primary, secondary, and tertiary amines, as well as cyclicamines and substituted amines such as naturally occurring andsynthesized substituted amines. Other pharmaceutically acceptableorganic non-toxic bases from which salts can be formed include ionexchange resins such as, for example, arginine, betaine, caffeine,choline, N′,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like.

When the compound of the present invention is basic, its correspondingsalt can be conveniently prepared from pharmaceutically acceptablenon-toxic acids, including inorganic and organic acids. Such acidsinclude, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic,citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic,hydrochloric, isethionic, lactic, maleic, malic, mandelic,methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric,succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like.Particularly preferred are citric, hydrobromic, hydrochloric, maleic,phosphoric, sulfuric, methanesulfonic, and tartaric acids.

The pharmaceutical compositions of the present invention or used by themethods of the present invention comprise a compound represented byFormula (I) (or a pharmaceutically acceptable salt or N-oxide thereof)as an active ingredient, a pharmaceutically acceptable carrier andoptionally other therapeutic ingredients or adjuvants. The compositionsinclude compositions suitable for oral, rectal, topical, and parenteral(including subcutaneous, intramuscular, and intravenous) administration,although the most suitable route in any given case will depend on theparticular host, and nature and severity of the conditions for which theactive ingredient is being administered. The pharmaceutical compositionsmay be conveniently presented in unit dosage form and prepared by any ofthe methods well known in the art of pharmacy.

In practice, the compounds represented by Formula (I), orpharmaceutically acceptable salts or N-oxides thereof, of this inventioncan be combined as the active ingredient in intimate admixture with apharmaceutical carrier according to conventional pharmaceuticalcompounding techniques. The carrier may take a wide variety of formsdepending on the form of preparation desired for administration. E.g.,oral or parenteral (including intravenous). Thus, the pharmaceuticalcompositions of the present invention can be presented as discrete unitssuitable for oral administration such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient.Further, the compositions can be presented as a powder, as granules, asa solution, as a suspension in an aqueous liquid, as a non-aqueousliquid, as an oil-in-water emulsion, or as a water-in-oil liquidemulsion. In addition to the common dosage forms set out above, thecompound represented by Formula (I), or a pharmaceutically acceptablesalt or N-oxide thereof, may also be administered by controlled releasemeans and/or delivery devices. The compositions may be prepared by anyof the methods of pharmacy. In general, such methods include a step ofbringing into association the active ingredient with the carrier thatconstitutes one or more necessary ingredients. In general, thecompositions are prepared by uniformly and intimately admixing theactive ingredient with liquid carriers or finely divided solid carriersor both. The product can then be conveniently shaped into the desiredpresentation.

Thus, the pharmaceutical compositions of this invention may include apharmaceutically acceptable carrier and a compound or a pharmaceuticallyacceptable salt or N-oxide of Formula (I). The compounds of Formula (I),or pharmaceutically acceptable salts or N-oxides thereof, can also beincluded in pharmaceutical compositions in combination with one or moreother therapeutically active compounds.

The pharmaceutical compositions of this invention include apharmaceutically acceptable liposomal formulation containing a compoundof Formula (I) or a pharmaceutically acceptable salt or N-oxide thereof.

The pharmaceutical carrier employed can be, for example, a solid,liquid, or gas. Examples of solid carriers include lactose, terra alba,sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, andstearic acid. Examples of liquid carriers are sugar syrup, peanut oil,olive oil, and water. Examples of gaseous carriers include carbondioxide and nitrogen.

In preparing the compositions for oral dosage form, any convenientpharmaceutical media may be employed. For example, water, glycols, oils,alcohols, flavoring agents, preservatives, coloring agents, and the likemay be used to form oral liquid preparations such as suspensions,elixirs and solutions; while carriers such as starches, sugars,microcrystalline cellulose, diluents, granulating agents, lubricants,binders, disintegrating agents, and the like may be used to form oralsolid preparations such as powders, capsules and tablets. Because oftheir ease of administration, tablets and capsules are the preferredoral dosage units whereby solid pharmaceutical carriers are employed.Optionally, tablets may be coated by standard aqueous or nonaqueoustechniques.

A tablet containing the composition of this invention may be prepared bycompression or molding, optionally with one or more accessoryingredients or adjuvants. Compressed tablets may be prepared bycompressing, in a suitable machine, the active ingredient in afree-flowing form such as powder or granules, optionally mixed with abinder, lubricant, inert diluent, surface active or dispersing agent orother such excipient. These excipients may be, for example, inertdiluents such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, corn starch, or alginic acid; binding agents, for example,starch, gelatin or acacia; and lubricating agents, for example,magnesium stearate, stearic acid or talc. The tablets may be uncoated orthey may be coated by known techniques to delay disintegration andabsorption in the gastrointestinal tract and thereby provide a sustainedaction over a longer time. For example, a time delay material such asglyceryl monostearate or glyceryl distearate may be used.

In hard gelatin capsules, the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin. In soft gelatin capsules, the active ingredient is mixed withwater or an oil medium, for example, peanut oil, liquid paraffin orolive oil. Molded tablets may be made by molding in a suitable machine,a mixture of the powdered compound moistened with an inert liquiddiluent. Each tablet preferably contains from about 0.05 mg to about 5 gof the active ingredient and each cachet or capsule preferablycontaining from about 0.05 mg to about 5 g of the active ingredient.

For example, a formulation intended for the oral administration tohumans may contain from about 0.5 mg to about 5 g of active agent,compounded with an appropriate and convenient amount of carriermaterial, which may vary from about 5 to about 95 percent of the totalcomposition. Unit dosage forms will generally contain between from about1 mg to about 2 g of the active ingredient, typically 25 mg, 50 mg, 100mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

Pharmaceutical compositions of the present invention suitable forparenteral administration may be prepared as solutions or suspensions ofthe active compounds in water. A suitable surfactant can be includedsuch as, for example, hydroxypropylcellulose. Dispersions can also beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofin oils. Further, a preservative can be included to prevent thedetrimental growth of microorganisms.

Pharmaceutical compositions of the present invention suitable forinjectable use include sterile aqueous solutions or dispersions.Furthermore, the compositions can be in the form of sterile powders forthe extemporaneous preparation of such sterile injectable solutions ordispersions. In all cases, the final injectable form must be sterile andmust be effectively fluid for easy syringability. The pharmaceuticalcompositions must be stable under the conditions of manufacture andstorage; thus, preferably should be preserved against the contaminatingaction of microorganisms such as bacteria and fungi. The carrier can bea solvent or dispersion medium containing, for example, water, ethanol,polyol (e.g., glycerol, propylene glycol and liquid polyethyleneglycol), vegetable oils, and suitable mixtures thereof.

Pharmaceutical compositions of the present invention can be in a formsuitable for topical use such as, for example, an aerosol, cream,ointment, lotion, dusting powder, or the like. Further, the compositionscan be in a form suitable for use in transdermal devices. Theseformulations may be prepared, utilizing a compound represented byFormula (I) of this invention, or a pharmaceutically acceptable salt orN-oxide thereof, via conventional processing methods. As an example, acream or ointment is prepared by admixing hydrophilic material andwater, together with about 5 wt % to about 10 wt % of the compound, toproduce a cream or ointment having a desired consistency.

Pharmaceutical compositions of this invention can be in a form suitablefor rectal administration wherein the carrier is a solid. It ispreferable that the mixture forms unit dose suppositories. Suitablecarriers include cocoa butter and other materials commonly used in theart. The suppositories may be conveniently formed by first admixing thecomposition with the softened or melted carrier(s) followed by chillingand shaping in molds.

In addition to the aforementioned carrier ingredients, thepharmaceutical formulations described above may include, as appropriate,one or more additional carrier ingredients such as diluents, buffers,flavoring agents, binders, surface-active agents, thickeners,lubricants, preservatives (including anti-oxidants) and the like.Furthermore, other adjuvants can be included to render the formulationisotonic with the blood of the intended recipient. Compositionscontaining a compound described by Formula (I), or pharmaceuticallyacceptable salts or N-oxides thereof, may also be prepared in powder orliquid concentrate form.

Generally, dosage levels on the order of from about 0.01 mg/kg to about150 mg/kg of body weight per day are useful in the treatment of theabove-indicated conditions, or alternatively about 0.5 mg to about 10 gper patient per day. For example, breast cancer, head and neck cancers,and gastrointestinal cancer such as colon, rectal or stomach cancer maybe effectively treated by the administration of from about 0.01 to 100mg of the compound per kilogram of body weight per day, or alternativelyabout 0.5 mg to about 7 g per patient per day.

Similarly, leukemia, ovarian, bronchial, lung, and pancreatic cancer maybe effectively treated by the administration of from about 0.01 to 100mg of the compound per kilogram of body weight per day, or alternativelyabout 0.5 mg to about 7 g per patient per day.

Mastocytosis/mast cell leukemia, gastrointestinal stromal tumors (GIST),small cell lung carcinoma (SCLC), sinonasal natural killer/T-celllymphoma, testicular cancer (seminoma), thyroid carcinoma, malignantmelanoma, ovarian carcinoma, adenoid cystic carcinoma, acute myelogenousleukemia (AML), breast carcinoma, pediatric T-cell acute lymphoblasticleukemia, angiosarcoma, anaplastic large cell lymphoma, endometrialcarcinoma, and prostate carcinoma may be effectively treated by theadministration of from about 0.01 to 100 mg of the compound per kilogramof body weight per day, or alternatively about 0.5 mg to about 7 g perpatient per day.

It is understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theage, body weight, general health, sex, diet, time of administration,route of administration, rate of excretion, drug combination and theseverity of the particular disease undergoing therapy.

The compounds of the present invention, or pharmaceutically acceptablesalts or N-oxides thereof, can also be effectively administered inconjunction with other cancer therapeutic compounds. For example,cytotoxic agents and angiogenesis inhibiting agents can be advantageousco-agents with the compounds of the present invention. Accordingly, thepresent invention includes compositions comprising the compoundsrepresented by Formula (I), or a pharmaceutically acceptable salt orN-oxide thereof, and a cytotoxic agent or an angiogenesis-inhibitingagent. The amounts of each can be therapeutically effective alone—inwhich case the additive effects can overcome cancers resistant totreatment by monotherapy. The amounts of any can also besubtherapeutic—to minimize adverse effects, particularly in sensitivepatients.

It is understood that the treatment of cancer depends on the type ofcancer. For example, lung cancer is treated differently as a first linetherapy than are colon cancer or breast cancer treated. Even within lungcancer, for example, first line therapy is different from second linetherapy, which in turn is different from third line therapy. Newlydiagnosed patients might be treated with cisplatinum containingregimens. Were that to fail, they move onto a second line therapy suchas a taxane. Finally, if that failed, they might get a tyrosine kinaseEGFR inhibitor as a third line therapy. Further, The regulatory approvalprocess differs from country to country. Accordingly, the acceptedtreatment regimens can differ from country to country. Nevertheless, thecompounds of the present invention, or pharmaceutically acceptable saltsor N-oxides thereof, can be beneficially co-administered in conjunctionor combination with other such cancer therapeutic compounds. Such othercompounds include, for example, a variety of cytotoxic agents(alkylators, DNA topoisomerase inhibitors, antimetabolites, tubulinbinders); inhibitors of angiogenesis; and different other forms oftherapies including kinase inhibitors such as Tarceva, monoclonalantibodies, and cancer vaccines. Other such compounds that can bebeneficially co-administered with the compounds of the present inventioninclude doxorubicin, vincristine, cisplatin, carboplatin, gemcitabine,and the taxanes. Thus, the compositions of the present invention includea compound according to Formula (I), or a pharmaceutically acceptablesalt or N-oxide thereof, and an anti-neoplastic, anti-tumor,anti-angiogenic, or chemotherapeutic agent.

The compounds of the present invention, or pharmaceutically acceptablesalts or N-oxides thereof, can also be effectively administered inconjunction with other therapeutic compounds, aside from cancer therapy.For example, therapeutic agents effective to ameliorate adverseside-effects can be advantageous co-agents with the compounds of thepresent invention.

C-KIT H526 Cell Assay Protocol

I. Assay for Inhibition of C-Kit in Intact Cells

The ability of compounds to inhibit the tyrosine kinase activity ofc-Kit was determined in a cell-based ELISA assay using the H526 cellline (ATCC # CRL-5811), which was originally derived from a human smallcell lung cancer. The assay determines the ability of compounds to blockligand-stimulated tyrosine phosphorylation of the wild-type c-Kitreceptor protein that is endogenously expressed in H526 cells. Cells arepre-incubated with compounds at various concentrations prior to additionof stem cell factor (SCF), the ligand for the c-Kit receptor tyrosinekinase. Cell lysates are then prepared and the c-Kit protein is capturedonto a c-Kit antibody-coated 96-well ELISA plate. The phosphotyrosinecontent of the receptor protein is then monitored by quantitation of thedegree of binding of an antibody that recognizes only the phosphorylatedtyrosine residues within the captured protein. The antibody used has areporter enzyme (e.g. horseradish peroxidase, HRP) covalently attached,such that binding of antibody to phosphorylated c-Kit can be determinedquantitatively by incubation with an appropriate HRP substrate.

In the assays below, the following abbreviations are used: HRP forhorseradish peroxidase, BSA for bovine serum albumin, EDTA forehtylenediaminetetraacetic acid, PBS for phosphate-buffered saline, SCFfor stem cell factor, DMSO for dimethylsulfoxide, rt for roomtemperature, min for minute, and h for hour.

The stock reagents used are as follows:

Cell lysis buffer:

50 mM Tris-HCl, pH 7.4

150 mM NaCl

10% Glycerol

1% Triton X-100

0.5 mM EDTA

1 μg/mL leupeptin

1 μg/mL aprotinin

1 mM sodium orthovanadate

Anti c-Kit antibody:

0.5 μg/mL anti c-Kit Ab-3 (Lab Vision, catalog #MS289P1) in 50 mM Sodiumbicarbonate, pH 9.

ELISA Assay Plates:

ELISA assay plates are prepared by addition of 100 μL of anti c-Kitantibody to each well of a 96-well Microlite-2 plate (Dynex, catalog #7417), followed by incubation at 37° C. for 2 h. The wells are thenwashed twice with 300 μL wash buffer.

Plate Wash Buffer:

PBS containing 0.5% Tween-20 (PBST)

Cell Assay Medium:

RPMI with 0.1% BSA

pY20-HRP:

25 ng/mL pY20-HRP (Calbiochem, catalog # 525320) in PBS, containing 0.5%Tween-20, 5% BSA, 1 mM Sodium orthovanadate

HRP Substrate:

Chemoluminescent detection reagent (Pierce, catalog # 37075)

Assay Protocol

Cultures of H526 cells, growing in RPMI with 10% fetal calf serum, werecollected by centrifugation, washed twice with PBS, and suspended incell assay medium. Cells were then distributed into a V-bottom 96-wellplate at 7.5×10⁴ cells per well in 100 μL cell assay medium.

Compound dilutions were prepared from 10 mM DMSO stocks by dilution incell assay medium, the final concentration of DMSO in the assay being0.1%. To compound incubation wells, 50 μL of the test compound was added(compounds are assayed at concentrations between 0.1 nM and 100 μM); topositive and negative control wells, 50 μL cell assay medium containing0.1% DMSO was added. The cells were then incubated with compound at 37°C. for 3 h. SCF (R&D Systems, catalog #255-SC-010) was then added inorder to stimulate the c-Kit receptor and induce its tyrosinephosphorylation. Then, 10 μL of a 1.6 μg/mL solution of SCF in cellassay medium was added to all wells apart from the negative controlwells, and the cells were incubated for an additional 15 min at 37° C.Following the addition of ice-cold PBS, the plate was centrifuged at1000 rpm for 5 min, the medium removed by aspiration, and the cellpellet lysed by the addition of 120 μL ice-cold cell lysis buffer perwell. The plate was kept on ice for 20 min and 100 μL of the celllysates from each well were then transferred to the wells of an ELISAassay plate and incubated at 4° C. for 16 h.

Following incubation of the cell lysates in the ELISA plate, the wellswere washed 4 times with 300 μL wash buffer, then 100 μL of thephosphotyrosine detection antibody pY20-HRP was added to each well andthe plate incubated at rt for 2 h. The wells were then washed 4 timeswith 300 μL wash buffer. Then, 50 μL of the chemiluminescent HRPsubstrate was added to each well for luminometric quantitation of theamount of antiphosphotyrosine-HRP conjugate bound to the plate.

Comparison of the assay signals obtained in the presence of compoundwith those of the positive and negative controls (cells incubated in thepresence or absence of SCF, with no compound added), allows the degreeof inhibition of c-Kit receptor tyrosine phosphorylation to bedetermined over a range of compound concentrations. These inhibitionvalues were fitted to a sigmoidal dose-response inhibition curve todetermine the IC50 values (i.e. the concentration of compound thatinhibits SCF-induced tyrosine phosphorylation of the c-Kit protein by50%).

The compounds of this invention reduced the ability of Kit tophosphorylate poly(Glu:Tyr) in the above assay, thus demonstratingdirect inhibition of the c-Kit receptor tyrosine kinase activity. IC₅₀values in this assay for the examples below were between 0.2 μM and 10μM. COMPOUNDS 1-2 (see experimental) had IC₅₀ values in this assaygreater than 50 μM.

EXPERIMENTAL

The EXAMPLES of the present invention were prepared according to thefollowing procedures:

Referring to the scheme shown below for EXAMPLE 1, reaction ofaminothiophene 1 with aldehydes under reducing conditions affordssecondary amines such as compound 2—for example, in the presence of amixture of triethylsilane and trifluoroacetic acid, or other reagentssuch as (but not limited to) sodium cyanoborohydride, sodiumtriacetoxyborohydride, sodium borohydride and hydrogen. Saponificationof the resulting ester then gives carboxylic acids of type 3.

Compounds such as 3 may then be reacted with hydrazines or alkoxaminesin the presence of an activating agent to give hydrazides such asEXAMPLE 1 or alkoxamides such as EXAMPLE 3.

In the section below, the following abbreviations are used: Me formethyl, Et for ethyl, Ph for phenyl, iPr for isopropyl, DMF fordimethylformamide, EDC for 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride, HOAt for 1-hydroxy-7-azabenzotriazole, EtOAc for ethylacetate, DMSO for dimethylsulfoxide, DCM for dichloromethane, TFA fortrifluoroacetic acid, MS for mass spectroscopy, ES for electrospray, rtfor room temperature, min for minute, and h for hour.

Example 1N′-(3,4-dichlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide

EXAMPLE 1 was prepared by the following procedure:

Part 1

Methyl 3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxylate (2): Amixture of 3-amino-thiophene-2-carboxylic acid methyl ester (5 g, 31.8mmol) and 4-quinoline-carboxaldehyde (5.25 g, 33.4 mmol) in TFA/CH₂Cl₂(75 mL, 75 mL) was heated at 50° C. for 3.5 h. The solution was cooledin an ice bath and triethylsilane (10.2 mL, 63.6 mmol) was addeddrop-wise over 5 min. The reaction mixture was stirred at 50° C. for 3.5h, cooled to rt, and 500 mL of CH₂Cl₂ was added. The reaction mixturewas basified with 10N NaOH (pH 6-7) followed by sat. NaHCO₃ (pH 8). TheCH₂Cl₂ layer was separated and the aqueous layer was extracted withCH₂Cl₂ (2×100 mL). The organic extracts were combined, washed withbrine, dried over anhydrous sodium sulfate, filtered, and concentratedin vacuo to yield the crude product, which was triturated with hexane togive pure methyl 3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxylateas white solid. MS (ES): m/z 298.55 (100) [MH⁺]; ¹H-NMR (400 MHz/CDCl₃):δ 3.87 (s, 3H), 5.00 (d, J=4.0 Hz, 2H), 6.48 (d, J=5.6 Hz, 1H), 7.30 (d,J=5.6 Hz, 1H), 7.36 (m, 1H), 7.41 (d, J=4.4 Hz, 1H), 7.62 (t, J=8.0 Hz,1H), 7.76 (t, J=9.6 Hz, 1H), 8.01 (d, J=8.0 Hz, 1H), 8.17 (d, J=8.4 Hz,1H), 8.86 (d, J=4.4 Hz, 1H).

Part 2

3-[(Quinolin-4-ylmethyl)amino]thiophene-2-carboxylic acid hydrochloride(3): To a suspension of3-[(quinolin-4-ylmethyl)-amino]-thiophene-2-carboxylic acid methyl ester(5.00 g, 16.8 mmoles) in a mixture of H₂O (20 mL) and MeOH (250 mL) wasadded solid NaOH (6.7 g, 167.5 mmoles). The reaction mixture was stirredat reflux for 5 h. The organic solvent was evaporated under reducedpressure and water (60 mL) was added to dissolve the residue. Theaqueous solution was washed with ethyl acetate (50 mL) and thenacidified with 1N HCl to pH 5-6.3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxylic acid hydrochlorideprecipitated from solution and was collected by filtration (3.86 g).¹HNMR (400 MHz/DMSO-d₆): δ 5.07 (s, 2H), 6.71 (d, J=4.0 Hz, 1H), 7.36(d, J=4.0 Hz, 1H), 7.43 (m, 1H), 7.58 (d, J=4.0 Hz, 1H), 7.67 (t, J=8.4Hz, 1H), 7.80 (t, J=7.2 Hz, 1H), 8.06 (d, J=8.4 Hz, 1H), 8.22 (d, J=8.4Hz, 1H), 8.83 (d, J=3.6 Hz, 1H), 12.30 (s, 1H). MS (ES⁺): 285[MH⁺].

Part 3

N′-(3,4-dichlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:To a solution of 3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxylicacid hydrochloride (100 mg, 0.31 mmol), EDC (192 mg, 1.00 mmol), HOAt(0.1 mL, 0.06 mmol, 0.5-0.7 M in DMF), and ^(i)Pr₂NEt (0.3 mL, 1.72mmol) in CH₂Cl₂ (7 mL) was added 3,4-dichlorophenylhydrazine (186 mg,1.05 mmol). The mixture was then stirred at rt. After the reaction wascomplete (˜16 h), saturated aqueous NaHCO₃ was added and layers wereseparated. The aqueous phase was extracted with CH₂Cl₂. The organicphases were combined, dried over Na₂SO₄, filtered, concentrated invacuo, and purified by silica gel chromatography to give EXAMPLE 1.¹H-NMR (400 MHz/CDCl₃): δ=4.98 (d, J=6.4 Hz, 2H), 6.13 (s, 1H), 6.53 (d,J=5.6 Hz, 1H), 6.79 (dd, J=4.0, 12 Hz, 1H), 7.05 (d, J=2.4 Hz, 1H), 7.31(d, J=8.8 Hz, 1H), 7.40 (d, J=4.4 Hz, 1H), 7.61 (t, J=8.0 Hz, 1H), 7.75(t, J=8.0 Hz, 1H), 8.0 (d, J=8.0 Hz, 1H), 8.17 (d, J=8.0 Hz, 1H), 8.85(d, J=4.4 Hz, 1H). MS (ES⁺): 443 [MH⁺].

The following analogues were prepared using3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxylic acid hydrochloride(EXAMPLE 1, part 2) and the appropriate hydrazine or alkoxamine,according to the procedure described above for EXAMPLE 1, part 3.

Example 2N′-(4-chlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 409 [MH+]

Example 3N-(tert-butoxy)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide:MS (ES+) 356 [MH+]

Example 4N-piperidin-1-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide:MS (ES+) 367 [MH+]

Example 5N′-(4-fluorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 393 [MH+]

Example 6N′-(4-bromophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 453 [MH+]

Example 7N-(4-methylpiperazin-1-yl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide:MS (ES+) 382 [MH+]

Example 8N-2,3-dihydro-1H-indol-1-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide:MS (ES+) 401 [MH+]

Example 9N-phenoxy-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide: MS(ES+) 376 [MH+]

Example 10N′-methyl-N′-phenyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 389 [MH+]

Example 11N′-(3-chlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 409 [MH+]

Example 12N′-phenyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide: MS(ES+) 375 [MH+]

Example 13N′-(4-cyanophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 400 [MH+]

Example 143-[(quinolin-4-ylmethyl)amino]-N′-[4-(trifluoromethyl)phenyl]thiophene-2-carbohydrazide:MS (ES+) 443 [MH+]

Example 15N′-(4-methoxyphenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 404 [MH+]

Example 163-[(quinolin-4-ylmethyl)amino]-N′-[3-(trifluoromethyl)phenyl]thiophene-2-carbohydrazide:MS (ES+) 443 [MH+]

Compound 1N′-(tert-butyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide:MS (ES+) 355 [MH+]

Compound 2N-morpholin-4-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide:MS (ES+) 369 [MH+]

1. A compound represented by Formula (I):

or a pharmaceutically acceptable salt or N-oxide thereof, wherein: J is—NR¹R² or —OR¹; A is aryl, heteroaryl, cycloC₃₋₁₀alkyl, heterocyclyl,cycloC₃₋₁₀alkenyl, or heterocycloalkenyl, each of which is optionallysubstituted by 1-5 independent R⁴ substituents; R¹ is C₁₋₆alkyl, aryl,heteroaryl, cycloC₃₋₁₀alkyl, heterocyclyl, cycloC₃₋₁₀alkenyl,heterocycloalkenyl, C₂₋₆alkenyl, C₂₋₆alkynyl, each of which isoptionally substituted by 1-5 independent R⁴¹ substituents; R² isC₀₋₆alkyl, aryl, heteroaryl, cycloC₃₋₁₀alkyl, heterocyclyl,cycloC₃₋₁₀alkenyl, heterocycloalkenyl, C₂₋₆alkenyl, C₂₋₆alkynyl, each ofwhich is optionally substituted by 1-5 independent R⁴² substituents; orR¹ and R², taken together with the N atom to which they are attached,form a heterocyclyl, heterocylcoalkenyl, or heteroaryl, each of which isoptionally substituted with 1-4 independent R⁴¹ substituents; providedR¹ and R² taken together with the N atom to which they are attached donot form a pyrazolyl; R³ is C₀₋₆alkyl, halogen, C₁₋₆alkyl, or haloalkyl;R⁴, R⁴¹, and R⁴² each independently is C₀₋₆alkyl, cycloC₃₋₁₀alkyl, oxo,halogen, haloalkyl, cyanoC₀₋₆alkyl, nitroC₀₋₆alkyl, hydroxyC₀₋₆alkyl,C₀₋₆alkylaminoC₀₋₆alkyl, aminoC₁₋₆alkylamino, acylaminoC₀₋₆alkylamino,acylC₀₋₆alkyl, substituted acyl, guanidinoC₀₋₆alkyl,hydroxyiminoC₀₋₆alkyl, acylaminoC₀₋₆alkyl, substituted acylamino,acyloxyC₀₋₆alkyl, substituted acyloxy, arC₀₋₆alkyl, substitutedarC₀₋₆alkyl, heteroarylC₀₋₆alkyl, substituted heteroarylC₀₋₆alkyl,heterocyclylC₀₋₆alkyl, cyanoaminoC₀₋₆alkyl, C₀₋₆alkylhydrazino,heterocyclylamino, arC₀₋₆alkylhydrazino, alkylsulfonylC₀₋₆alkyl,arC₀₋₆alkylsulfonylC₀₋₆alkyl, alkylsulfinylC₀₋₆alkyl,alkylsulfonamidoC₀₋₆alkyl, arC₀₋₆alkylsulfonamidoC₀₋₆alkyl,aminoC₀₋₆alkylsulfonyl, C₀₋₆alkylaminosulfonyl, acylC₁₋₆alkylsulfonyl,heterocyclylsulfonyl, aminoC₀₋₆alkylsulfinyl, acylC₁₋₆alkylsulfinyl,silyl, siloxy, alkenoxy, alkynoxy, C₂₋₆alkenyl, acylC₂₋₆alkenyl,C₂₋₆alkynyl, acylC₂₋₆alkynyl, hydroxyC₂₋₆alkynyl, aminoC₂₋₆alkynyl,C₁₋₆alkoxyC₀₋₆alkyl, C₁₋₆alkylthioC₀₋₆alkyl, hydroxyC₁₋₆alkoxyC₀₋₆alkyl,hydroxyC₁₋₆alkylthioC₀₋₆alkyl, acylC₁₋₆alkoxyC₀₋₆alkyl,acylC₁₋₆alkylthioC₀₋₆alkyl, C₀₋₆alkylaminoC₁₋₆alkoxyC₀₋₆alkyl,C₀₋₆alkylaminoC₁₋₆alkylthioC₀₋₆alkyl, acylaminoC₁₋₆alkoxyC₀₋₆alkyl,acylaminoC₁₋₆alkylthioC₀₋₆alkyl, arC₀₋₆alkylaminoC₀₋₆alkyl,arC₀₋₆alkylthioC₀₋₆alkyl, arC₀₋₆alkoxyC₀₋₆alkyl, arC₀₋₆alkylamino,arC₀₋₆alkylaminoC₀₋₆alkyl, arC₀₋₆alkylthio, substituted arC₀₋₆alkoxy,substituted arC₀₋₆alkylthio,or substituted arC₀₋₆alkoxy; and providedthat the compound is notN′-(tert-butyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazideorN-morpholin-4-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide.2. The compound according to claim 1 wherein J is —NR¹R².
 3. Thecompound according to claim 2 wherein R³ is hydrogen.
 4. The compoundaccording to claim 2 wherein A is heteroaryl.
 5. The compound accordingto claim 2 wherein A is


6. The compound according to claim 1 wherein J is —OR¹.
 7. The compoundaccording to claim 6 wherein R³ is hydrogen.
 8. The compound accordingto claim 6 wherein A is heteroaryl.
 9. The compound according to claim 6wherein A is


10. A composition comprising a compound according to claim 1, or apharmaceutically acceptable salt or N-oxide thereof, and apharmaceutically acceptable carrier.
 11. A composition comprising acompound according to claim 1, or a pharmaceutically acceptable salt orN-oxide thereof; and an anti-neoplastic, anti-tumor, anti-angiogenic, orchemotherapeutic agent.
 12. A composition comprising a compoundaccording to claim 1, or a pharmaceutically acceptable salt or N-oxidethereof, and a cytotoxic cancer therapeutic agent.
 13. A compositioncomprising a compound according to claim 1, or a pharmaceuticallyacceptable salt or N-oxide thereof, and an angiogenesis inhibitingcancer therapeutic agent.
 14. A compound consisting ofN′-(4-chlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N-(tert-butoxy)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;N-piperidin-1-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;N′-(4-bromophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N-(4-methylpiperazin-1-yl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;N′-(3,4-dichlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N-phenoxy-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamide;N′-methyl-N′-phenyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N′-(3-chlorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N′-phenyl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N′-(4-cyanophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;3-[(quinolin-4-ylmethyl)amino]-N′-[4-(trifluoromethyl)phenyl]thiophene-2-carbohydrazide;N′-(4-methoxyphenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;3-[(quinolin-4-ylmethyl)amino]-N′-[3-(trifluoromethyl)phenyl]thiophene-2-carbohydrazide;N′-(4-fluorophenyl)-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carbohydrazide;N-2,3-dihydro-1H-indol-1-yl-3-[(quinolin-4-ylmethyl)amino]thiophene-2-carboxamideor a pharmaceutically acceptable salt, or N-oxide, thereof.